Doxycycline Formulations, and Methods of Treating Rosacea

ABSTRACT

The present invention is directed to a pharmaceutical composition in unit dose form for orally delivering doxycycline to a human, the pharmaceutical composition comprising: a capsule, wherein the capsule is coated with a delayed release layer; wherein the delayed release layer comprises about 4 to 6 mg of doxycycline monohydrate and a binding agent, and wherein the delayed release layer is coated with an enteric coating; wherein the enteric coating dissolves at pH of about 5 to 6, and wherein the enteric coating is coated with an immediate release layer; wherein the immediate release layer comprises about 32 mg of doxycycline monohydrate and a binding agent, wherein the relative mean C max  of the pharmaceutical composition is within 80.00% to 125.00% of a C max  value of 510±220.7 ng/mL, after administration of a single dose of the pharmaceutical composition to humans in a fasting state; and wherein the relative mean AUC (0-∞)  of the pharmaceutical composition is within 80.00% to 125.00% of a AUC (0-∞)  value of 9227±3212.8 ng·hr/mL, after administration of a single dose of the pharmaceutical composition to humans in a fasting state.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.14/711,375, filed May 13, 2015, which is a continuation of U.S.application Ser. No. 14/055,659, filed Oct. 16, 2013, which is acontinuation of U.S. application Ser. No. 13/929,598, filed Jun. 27,2013, now U.S. Pat. No. 8,652,516, which claims the benefit of U.S.Provisional Application Ser. No. 61/794,030, filed Mar. 15, 2013. Allaforementioned applications are incorporated herein by reference intheir entireties.

BACKGROUND OF THE INVENTION

Rosacea is a chronic condition characterized by facial erythema and skinlesions. Rosacea typically begins as redness on the central face acrossthe cheeks, nose, or forehead, but can also less commonly affect theneck, chest, ears, and scalp. Symptoms, such as semi-permanent redness,telangiectasia, red domed papules and pustules, and in some advancedcases, a red lobulated nose (rhinophyma), may develop.

Although there are effective treatments for rosacea on the market (e.g.,Oracea® sold by Galderma Laboratories, L.P.), formulations with reducedactive ingredients are desirable.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graph showing the Mean (±SD) Plasma DoxycyclineConcentration vs. Time Profiles using Normal scale.

FIG. 2 is a graph showing the Mean (±SD) Plasma DoxycyclineConcentration vs. Time Profiles in Log-linear Scale.

FIG. 3 is a graph showing the Mean (±SD) Plasma DoxycyclineConcentration vs. Time Profiles using Normal scale.

FIG. 4 is a graph showing the Mean (±SD) Plasma DoxycyclineConcentration vs. Time Profiles in Log-linear Scale.

FIG. 5 is a flow chart outlining a manufacturing process forcompositions of the present invention.

FIG. 6 is a graph showing Dissolution profiles of Doxycycline coatedcapsules, 40 mg versus Oracea® capsules, 40 mg.

FIG. 7 is a graph showing Dissolution profiles of Capsules coated with 4mg and 6 mg doxycycline: Delayed release layer.

FIG. 8 is a graph showing Dissolution profiles of Capsules coated with36 mg and 38 mg doxycycline: Finished Drug Product.

FIG. 9 is a graph showing a Pharmacokinetic study of Doxycycline coatedcapsules 40 mg versus Oracea® capsules, 40 mg.

FIG. 10 is a graph showing a Pharmacokinetic study of Doxycycline coatedcapsules 36 mg versus Oracea® capsules, 40 mg.

FIG. 11 is a graph showing a Pharmacokinetic study of Doxycycline coatedcapsules 37.5 mg versus Oracea® capsules, 40 mg.

SUMMARY OF THE INVENTION

In one embodiment, the present invention provides a pharmaceuticalcomposition in unit dose form for orally delivering doxycycline to ahuman. The pharmaceutical composition comprises a coated capsule. Thecapsule is coated with a delayed release layer, enteric coating and animmediate release layer. The delayed release layer comprises about 4 to6 mg of doxycycline, or a pharmaceutically acceptable salt thereof, anda binding agent. The delayed release layer is coated with an entericcoating. The enteric coating dissolves at pH of about 5 to 6. Theenteric coating is coated with an immediate release layer. The immediaterelease layer comprises about 32 mg of doxycycline, or apharmaceutically acceptable salt thereof, and a binding agent.

In one embodiment, the mean C_(max) of the pharmaceutical composition iswithin about 80.00% to 125.00% of a C_(max) value of about 510±220.7ng/mL (or about 80% to 125% of a C_(max) value in the range of about 290ng/mL to 730 ng/mL), after administration of a single dose of thepharmaceutical composition to humans in a fasting state. The meanAUC_((0-∞)) of the pharmaceutical composition is within about 80.00% to125.00% of a AUC_((0-∞)) value of about 9227±3212.8 ng·hr/mL (or about80% to 125% of a AUC_((0-∞)) value in the range of about 6010 ng·hr/mLto 12,440 ng·hr/mL), after administration of a single dose of thepharmaceutical composition to humans in a fasting state.

The mean C_(max) of the pharmaceutical composition at steady state iswithin about 80.00% to 125.00% of a C_(max) value of about 600±194.2ng/mL (or about 80% to 125% of a C_(max) value in the range of about 405ng/mL to 795 ng/mL), after administration to humans in a fasting state,and the relative mean AUC_((0-t)) of the pharmaceutical composition atsteady state is within about 80.00% to 125.00% of a AUC_((0-t)) value ofabout 7543±2443.9 ng·hr/mL (or about 80% to 125% of a AUC_((0-t)) valuein the range of about 5090 ng·hr/mL to 9990 ng·hr/mL), afteradministration to humans in a fasting state.

Preferably, the doxycycline monohydrate is micronized. In oneembodiment, the micronized doxycycline monohydrate has less than about10 μm distribution for at least about 90% of particles.

In one embodiment, the pharmaceutical composition further comprises aseal coating between the capsule and the delayed release coating. In oneembodiment, the seal coating comprises a pH dependent ingredient,wherein the pH dependent ingredient dissolves at pH of about 5 to 6.

The capsule contains about 200 mg to about 260 mg of at least one inertingredient. Preferably, the capsule does not contain an activeingredient. Preferably, the capsule is hydroxypropyl methylcellulose.

The ratio of the binding agent to the doxycycline monohydrate in thedelayed release layer is about 1:3. The ratio of the binding agent tothe doxycycline monohydrate in the immediate release layer is about 1:3.Preferably, the binding agent is hydroxypropyl methylcellulose (HPMC),hydroxyethyl cellulose (HEC), hydroxypropyl cellulose (HPC), carboxymethyl cellulose (CMC), or mixtures thereof. More preferably, thebinding agent is hydroxypropyl methylcellulose (HPMC).

The pharmaceutical composition has a dissolution profile such that about80% to about 90% of the doxycycline monohydrate is dissolved after about30 minutes at pH of about 1.1. The pharmaceutical composition has adissolution profile such that about 90% of the doxycycline monohydrateis dissolved after about 120 minutes at pH of about 1.1 and 120 minutesat pH of about 6.0.

In one embodiment, the pharmaceutical composition further comprises anouter color coating.

In another embodiment, the present invention provides a method fortreating papules and pustules of rosacea in a human in need thereof,comprising orally administering a once a day dose of a pharmaceuticalcompositions of the present invention to the human.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is directed to pharmaceutical compositions thatproduce a sustained release profile of doxycycline, and methods of usingthe pharmaceutical compositions to treat disorders, in particularrosacea.

Throughout this specification, quantities are defined by ranges, and bylower and upper boundaries of ranges. Each lower boundary can becombined with each upper boundary to define a range. The lower and upperboundaries should each be taken as a separate element.

The pharmaceutical compositions of the present invention are preferablyprovided in unit dose form and are formulated for oral delivery ofdoxycycline to humans. In one embodiment, the unit dose form of thepharmaceutical composition comprises a hard or soft capsule containing afill. The fill consists of, or consists essentially of, one or moreinert ingredients in a pharmaceutically acceptable vehicle. The capsuleis coated by at least two separate layers comprising doxycycline.

The manufacture of hard or soft capsules is generally known by those ofordinary skill in the art. For example, soft capsules may be made byvarious processes including the plate process, the rotary die process,the reciprocating die process, and the continuous process. Examples ofthe capsular materials include, but are not limited to, natural orsynthetic gelatin, pectin, casein, collagen, protein, modified starch,polyvinyl pyrrolidone, acrylic polymers, cellulose derivatives (such as,but not limited to, hydroxypropyl methylcellulose (HPMC)), andcombinations thereof, optionally with one or more plasticizers and/orwater. Capsular materials may also include one or more preservatives,coloring and opacifying agents, flavorings and sweeteners, sugars,gastroresistant substances, or combinations thereof.

The shape and size of the capsules can vary. The shape of the capsulemay be, but is not limited to, round, oval, tubular, oblong, twist off,or a non-standard shape, preferably oblong. The size of the capsule usedwill vary in accordance to the volume of the fill composition intendedto be contained therein. The various standard sizes of capsules areconventionally designated as (000), (00), (0), (1), (2), (3), (4), and(5).

The preferred material for a capsule of the present invention is onethat retains doxycycline, or a pharmaceutically acceptable salt thereof,well and is not sensitive to temperature. An example of such is a HPMCcapsule. Also, the preferred capsule has an oval shape and is of sizedesignation (2).

The capsules are filled with pharmaceutically acceptable inertingredients. These ingredients typically function to provide weight tothe capsules so they do not fly around during coating processes. Theterm “inert ingredient” refers to any compound or compounds which do nothave, or substantially do not have, pharmacological or biologicalactivity. The capsules preferably do not contain any active ingredients.The capsules of the present invention are typically filled with about100 to about 300 mg, more typically, about 150 to about 250 mg, and mosttypically about 200 mg of inert ingredients.

Examples of inert ingredients include excipients, diluents,disintegrants, surfactants, lubricants, and glidants. Preferred inertingredients in the present invention include pregelatinized starch,lactose monohydrate and microcrystalline cellulose. A preferredlubricant is magnesium stearate.

The capsule of the present invention is coated with at least threelayers. In particular, the capsule is coated with a delayed releaselayer, which is coated with an enteric coating, which is coated with animmediate release layer.

The delayed release layer and the immediate release layer containdoxycycline. Doxycycline can be in the form of a pharmaceuticallyacceptable salt. The term “pharmaceutically acceptable salt” refers to asalt prepared from doxycycline and pharmaceutically acceptable non-toxicacids or bases. The acids may be inorganic or organic acids ofdoxycycline. Examples of inorganic acids include hydrochloric,hydrobromic, hydroiodic, sulfuric, and phosphoric acids. Examples oforganic acids include carboxylic and sulfonic acids. The radical of theorganic acids may be aliphatic or aromatic. Some examples of organicacids include formic, acetic, phenylacetic, propionic, succinic,glycolic, glucuronic, maleic, furoic, glutamic, benzoic, anthranilic,salicylic, phenylacetic, mandelic, embonic (pamoic), methanesulfonic,ethanesulfonic, panthenoic, benzenesulfonic, stearic, sulfanilic,alginic, tartaric, citric, gluconic, gulonic, arylsulfonic, andgalacturonic acids. Appropriate organic bases may be selected, forexample, from N,N-dibenzylethylenediamine, chloroprocaine, choline,diethanolamine, ethylenediamine, meglumine (N-methylglucamine), andprocaine.

Doxycycline is preferably administered as its hyclate salt or as ahydrate, preferably monohydrate.

The delayed release layer comprises doxycycline, or a salt thereof, anda binding agent. Examples of binding agents include cellulose polymers,such as, various grades of microcrystalline cellulose, hydroxypropylmethylcellulose (HPMC), hydroxyethyl cellulose (HEC), hydroxypropylcellulose (HPC), carboxy methyl cellulose (CMC), sodiumcarboxymethylcellulose, and mixtures thereof. Hydroxypropylmethylcellulose (HPMC) is preferred as the binding agent in the presentinvention.

In one embodiment, the delayed release layer comprises about 4 mg of adoxycycline, or a pharmaceutically acceptable salt thereof (e.g.,doxycycline monohydrate). In another embodiment, the delayed releaselayer comprises about 6 mg of a doxycycline, or a pharmaceuticallyacceptable salt thereof (e.g., doxycycline monohydrate). Typically, theratio of a binding agent to a doxycycline salt is about 1:3.

The delayed release layer may further comprise anti-foaming agents, suchas, for example, a simethicone emulsion. Typically, the ratio of abinding agent and anti-foaming agent to a doxycycline salt is about 1:3.

The delayed release layer is coated with an enteric coating. The entericcoating dissolves at pH of about 5 to 6, preferably at pH 5.5. Anexample of a preferred enteric coating is an aqueous dispersion ofanionic polymers with methacrylic acid as a functional group. Forinstance, Eudragit® L30-55 can be used in the enteric coating.Additionally, the enteric coating may further comprise plasticizers. Anexample of a preferred plasticizer is triethylcitrate.

The enteric coating is coated with an immediate release layer. Theimmediate release layer comprises about 32 mg of a doxycycline, or apharmaceutically acceptable salt thereof (e.g., doxycyclinemonohydrate), and a binding agent. Typically, the ratio of a bindingagent to a doxycycline salt is about 1:3.

The immediate release layer may further comprise anti-foaming agents,such as, for example, a simethicone emulsion. Typically, the ratio of abinding agent and anti-foaming agent to a doxycycline salt is about 1:3.

In a preferred embodiment, the doxycycline salt of the present inventionis micronized. For example, micronized doxycycline monohydrate can haveless than about 10 μm distribution for at least about 70% of particles,for at least about 80% of particles, more typically for at least about90% of particles, and most typically, for at least about 95% ofparticles.

In some embodiments, the pharmaceutical composition further comprises aseal coating between the capsule and the delayed release coating. Theseal coating functions to seal and prepare the capsules for subsequentcoating. The seal coating can be pH independent or pH dependent. Apreferred example of a pH independent coating is HPMC. An example of apH dependent seal coating is an aqueous dispersion of anionic polymerswith methacrylic acid as a functional group, e.g., Eudragit L30-55. Boththese seal coatings adhere well to the capsules and provide acceptablesurfaces for further coating; however, Eudragit L30-55 protects thecapsule longer in an acidic environment.

In some embodiments, the pharmaceutical composition further comprises anouter dye coating which provides the dosage unit with a color coating.

In some embodiments, the pharmaceutical compositions of the presentinvention have a dissolution profile such that about 80% to about 90% ofthe doxycycline salt is dissolved after about 30 minutes at pH of about1.1. Also, the pharmaceutical compositions have a dissolution profilesuch that about 90% of the doxycycline salt is dissolved after about 120minutes at pH of about 1.1 and 120 minutes at pH of about 6.0.

The present invention includes a method for treating the inflammatorylesions of rosacea in a human in need thereof. Inflammatory lesions ofrosacea include papules and pustules of rosacea. The method comprisesorally administering a once a day dose of a pharmaceutical compositionof the present invention. That is, a unit dose form (i.e., a capsule) isadministered daily.

The unit dose forms of the present invention are bioequivalent to areference formulation. The reference formulation is a gelatin capsulefilled with beads of doxycycline monohydrate, wherein the beads consistof 30 mg of immediate release beads and 10 mg of delayed release beads.For example, the reference formulation is a product for the treatment ofthe papules and pustules of rosacea sold by Galderma Laboratories, L.P.of Fort Worth, Tex. under the trademark Oracea®.

Bioequivalence herein is defined by the regulations of the United StatesFederal and Drug Administration. In particular, bioequivalence is foundif the 90% Confidence Interval of the relative mean C_(max), AUC_((0-t))and AUC_((0-∞)) of a pharmaceutical composition to the referenceformulation is within 80.00% to 125.00% upon administration to humans ina fasting state and in a fed state, in a noncompartmental analysis withlog transformed data.

An example of a fasting state is at least a ten hour fast prior toadministration of a pharmaceutical composition.

The relative mean C_(max) of a unit dosage form of the present inventionis within 80.00% to 125.00% of the C_(max) value of 510±220.7 ng/mL ofOracea®, after administration of a single dose to humans in a fastingstate.

The relative mean AUC_((0-∞)) of a unit dosage form of the presentinvention is within 80.00% to 125.00% of the AUC_((0-∞)) value of9227±3212.8 ng·hr/mL of Oracea®, after administration of a single doseto humans in a fasting state.

The relative mean C_(max) of a unit dosage form of the present inventionat steady state is within 80.00% to 125.00% of the C_(max) value of600±194.2 ng/mL of Oracea®, after administration to humans in a fastingstate.

The relative mean AUC_((0-t)) of a unit dosage form of the presentinvention at steady state is within 80.00% to 125.00% of the AUC_((0-t))value of 7543±2443.9 ng·hr/mL of Oracea®, after administration to humansin a fasting state.

Under fed conditions, C_(max) and AUC Oracea® decrease about 30-50% andabout 20-40%, respectively, or about 45% and about 25%, respectively.The C_(max) and AUC of the unit dosage form of the present inventiondecrease commensurately under fed conditions.

In one embodiment, the C_(max) of a unit dosage form of the presentinvention, after administration of a single dose to humans in a fastingstate, has a range of about 200 ng/mL to about 1120 ng/mL. Examples ofother preferred lower boundaries of this range include about 230, 250,300, 350, 400, 450 and 500 ng/mL. Examples of other preferred upperboundaries of this range include about 500, 550, 600, 650, 700, 750,800, 850, 900 and 950 ng/mL.

In one embodiment, the AUC_((0-∞)) of a unit dosage form of the presentinvention, after administration of a single dose to humans in a fastingstate, has a range of about 4800 ng·hr/mL to about 15,600 ng·hr/mL.Examples of other preferred lower boundaries of this range include about5000, 5500, 6500, 7500, 8500, 9500 and 10,000 ng·hr/mL. Examples ofother preferred upper boundaries of this range include about 10,000,11,000, 12,000, 13,000, 14,000 and 15,000 ng·hr/mL.

In one embodiment, the C_(max) of a unit dosage form of the presentinvention at steady state, after administration to humans in a fastingstate, has a range of about 320 ng/mL to about 1000 ng/mL. Examples ofother preferred lower boundaries of this range include about 350, 400,450, 500, 550, 600 and 660 ng/mL. Examples of other preferred upperboundaries of this range include about 660, 750, 800, 850, 900 and 950ng/mL.

In one embodiment, the AUC_((0-t)) of a unit dosage form of the presentinvention, at steady state, after administration to humans in a fastingstate, has a range of about 3500 ng·hr/mL to about 12,500 ng·hr/mL.Examples of other preferred lower boundaries of this range include about4000, 4500, 5500, 6500, 7500 and 8500 ng·hr/mL. Examples of otherpreferred upper boundaries of this range include about 8500, 9500,10,500 and 11,500 ng·hr/mL.

Under fed conditions, C_(max) and AUC of the unit dosage form of thepresent invention decrease about 30-50% and about 20-40%, respectively,or about 45% and about 25%, respectively.

The plasma concentrations of doxycycline achieved upon administration ofunit dose forms of the present invention are less than the concentrationrequired to treat bacterial diseases. Administration of the compositionsfor up to eighteen months demonstrates no detectable long term effectson bacterial flora of the oral cavity, skin, intestinal tract and/orvagina. For example, administration of a daily unit dose results in noreduction of skin microflora during a six-month treatment.

Preferably, upon a once-daily oral dosage of the pharmaceuticalcomposition, a human is provided with blood levels of the doxycycline ofabout 0.3 μg/ml to about 0.8 μg/ml.

In one embodiment, the present invention also provides methods oftreating acne. The method comprises orally administering a once a daydose of the pharmaceutical compositions of the present invention, i.e.,a daily dose of a unit dose form. As used herein, the term “acne” is adisorder of the skin characterized by papules, pustules, cysts, nodules,comedones, and other blemishes or skin lesions. These blemishes andlesions are often accompanied by inflammation of the skin glands andpilosebaceous follicles, as well as, microbial, especially bacterial,infection.

For the purposes of this specification, acne includes all known types ofacne. Some types of acne include, for example, acne vulgaris, cysticacne, acne atrophica, bromide acne, chlorine acne, acne conglobata, acnecosmetica, acne detergicans, epidemic acne, acne estivalis, acnefulminans, halogen acne, acne indurata, iodide acne, acne keloid, acnemechanica, acne papulosa, pomade acne, premenstral acne, acne pustulosa,acne scorbutica, acne scrofulosorum, acne urticata, acne varioliformis,acne venenata, propionic acne, acne excoriee, gram negative acne,steroid acne, and nodulocystic acne.

The pharmaceutical compositions of the present invention can be made aswould be known by a skilled artisan. Examples of making the compositionscan be found in U.S. patent application Ser. No. 12/466,261,incorporated herein by reference in its entirety.

EXAMPLES Example 1 A Randomized, Single-Center, Single-Dose, Open-Label,Three-Way Crossover, Pivotal Bioequivalence Study of Doxycycline 36 mgCapsules and Doxycycline 38 mg Capsules Compared with ORACEA®(Doxycycline 40 mg) Capsules in Healthy Human Subjects Under FastingConditions

The aim of the study was to compare the bioavailability of doxycycline“drug coated” capsules 36-mg and doxycycline “drug coated” capsules38-mg of the present invention, relative to the reference product, theFDA approved 40-mg Oracea® capsule under fasting conditions. The studyshowed that the bioavailability of Doxycycline “drug coated” Capsules36-mg and Doxycycline “drug coated” Capsules 38-mg, are bioequivalent tothe reference product, the FDA approved 40-mg Oracea® capsule underfasting conditions.

Study Objectives

Pharmacokinetics objectives: Comparison of the rate and extent ofabsorption and exposure of two doses of doxycycline monohydrate capsules(Test products) with Oracea® 40-mg capsules (Reference product) inhealthy subjects in order to determine bioequivalence under fastingconditions.

Safety objectives: tolerability and safety of single oraladministrations of three dose levels of doxycycline capsules in healthysubjects under fasting conditions.

Study Design

The study was conducted according to an open-label, randomized,three-period crossover design with a wash-out phase of 7 days minimum.Subjects received a single dose of the test formulations (Treatment A,36-mg capsule and treatment B, 38-mg capsule) and the reference(Treatment C, 40-mg capsule) products 30 minutes after an overnight fast(i.e. at least 10 hours) from food followed by a fast from food for atleast 4 hours post-dose.

Summary of Study Results Study Characteristics

The study was carried out in 44 healthy volunteers; however 5 subjectsmissed at least one treatment period, and these subjects were notconsidered for the bioequivalence assessment. Subject reasons formissing at least one treatment period include 2 cases of scheduleconflicts, 2 for personal reasons and 1 Adverse Event of tonsillitis(unrelated to study drug). The study protocol was designed to ensurethat a minimum of 38 subjects completed, and this was objective wasachieved with 39 evaluable subjects.

Bioequivalence Assessment:

Blood samples for pharmacokinetic profiling were taken up to 96 hourspost-dose, and doxycycline plasma concentrations were determined with avalidated LC-MS/MS method, LOQ: 15 ng/mL.

The pharmacokinetic (PK) parameters for doxycycline were determinedusing a standard non-compartmental method, and C_(max), AUC_(0-t) (witht being the last quantifiable point), AUC_(0-inf) were analyzedstatistically using log-transformed data. Bioequivalence was assessed byexamining the 90% confidence intervals for the ratio of the testformulation mean relative to the reference formulation mean.

The mean and standard deviations for the PK parameters are provided inTables 1 and 2 below, and the mean plasma profiles in normal scale andlog scale are shown in FIGS. 1 and 2. The statistical analyses of PKresults are summarized in Tables 3 and 4.

36-Mg Doxycycline Capsule Vs. 40 mg Oracea Capsule:

The calculated 90% confidence intervals of the mean test/referenceratios of C_(max), AUC_(0-t), AUC_(0-inf) were [92.73; 102.21], [87.03;93.03] and [89.15; 97.75], respectively. Of note, the FDA guidance(Bioavailability and Bioequivalence Studies for Orally Administered DrugProducts—General Considerations—March 2003) states that to be inside theacceptance interval, values should be at least 80.00 and no more than125.00. Therefore, all tested PK parameters (C_(max), AUC_(0-t),AUC_(0-inf)) were strictly within the bioequivalence range of80.00-125.00%.

38-Mg Doxycycline Capsule Vs. 40 mg Oracea Capsule:

The calculated 90% confidence intervals of the mean test/referenceratios of C_(max), AUC_(0-t), AUC_(0-inf) were [88.19; 98.47], [87.67;94.28] and [86.80; 94.47], respectively. Therefore, all tested PKparameters (C_(max), AUC_(0-t), AUC_(0-inf)) were strictly within thebioequivalence range of 80.00-125.00%.

In conclusion, Doxycycline 36-mg capsule and 40-mg Oracea® capsule andDoxycycline 38-mg capsule and 40-mg Oracea® under fasting conditions areconsidered bioequivalent with respect to C_(max), AUC_(0-t) andAUC_(0-inf) using the 80-125% FDA required criteria. Adverse eventsexperienced were transient and mild to moderate in severity; they werealso consistent with the known safety/tolerability profile ofdoxycycline.

TABLE 1 Overall pharmacokinetics parameters (Arithmetic mean ± SD) 36-mgDoxycycline 40-mg Oracea ® Capsule Capsule Parameters N (TEST) N(REFERENCE) AUC_(0-t) 39 39 (ng · hr/mL) Mean ± SD 6397 ± 1457 7101 ±1451 Min-Max 3936-10598 3856-10418 CV (%) 23% 20% AUC_(0-inf) 22 31 (ng· hr/mL) Mean ± SD 7151 ± 1184 7529 ± 1462 Min-Max 4875-9673  4437-11001CV (%) 17% 19% C_(max) (ng/mL) 39 39 Mean ± SD 504 ± 189 514 ± 176Min-Max 258-1113 251-1056 CV (%) 38% 34% AUC: area under theconcentration time curve; C_(max): observed peak drug concentration;Note: AUC_(0-inf) were calculated only for profiles meeting acceptancecriteria in linear regression.

TABLE 2 Overall pharmacokinetics parameters (Arithmetic mean ± SD) 38-mgDoxycycline 40-mg Oracea ® Capsule Capsule Parameters N (TEST) N(REFERENCE) AUC_(0-t) 39 39 (ng · hr/mL) Mean ± SD 6500 ± 1523 7101 ±1451 Min-Max 3488-10345 3856-10418 CV (%) 23% 20% AUC_(0-inf) 29 31 (ng· hr/mL) Mean ± SD 7011 ± 1605 7529 ± 1462 Min-Max 3962-10688 4437-11001CV (%) 23% 19% C_(max) (ng/mL) 39 39 Mean ± SD 489 ± 199 514 ± 176Min-Max 212-1145 251-1056 CV (%) 41% 34% AUC: area under theconcentration time curve; C_(max): observed peak drug concentration;Note: AUC_(0-inf) were calculated only for profiles meeting acceptancecriteria in linear regression.

TABLE 3 Summary of statistical analysis of pharmacokinetic data (36-mgv. 40-mg; N = 39) Least Square Mean Geometric Mean^(a) 90% ConfidenceInterval 36 mg 40 mg 36 mg 40 mg Lower Upper Doxycycline Oracea ®Doxycycline Oracea ® Ratio Limit Limit Capsule Capsule Capsule Capsule(%) (%) (%) AUC_(0-t) 8.744 8.849 6271.73 6970.06 89.98 87.03 93.03 (ng*h/mL) AUC_(0-inf) 8.850 8.919 6972.21 7468.95 93.35 89.15 97.75 (ng*h/mL) C_(max) 6.166 6.193 476.44 489.38 97.35 92.73 102.21 (ng/mL)^(a)Geometric means are based on least square means of ln-transformedvalues N: Number of subjects included in PK analysis.

TABLE 4 Summary of statistical analysis of pharmacokinetic data (38-mgv. 40 mg; N = 39) Least Square Mean Geometric Mean^(a) 90% ConfidenceInterval 38 mg 40 mg 38 mg 40 mg Lower Upper Doxycycline Oracea ®Doxycycline Oracea ® Ratio Limit Limit Capsule Capsule Capsule Capsule(%) (%) (%) AUC_(0-t) 8.754 8.849 6336.27 6969.69 90.91 87.67 94.28 (ng*h/mL) AUC_(0-inf) 8.820 8.919 6769.67 7475.68 90.56 86.80 94.47 (ng*h/mL) C_(max) 6.123 6.194 456.26 489.62 93.19 88.19 98.47 (ng/mL)^(a)Geometric means are based on least square means of ln-transformedvalues N: Number of subjects included in PK analysis

Example 2 A Randomized, Single-Center, Single-Dose, Open-Label,Three-Way Crossover, Pivotal Bioequivalence Study of Doxycycline 36 mgCapsules and Doxycycline 38 mg Capsules Compared with ORACEA®(Doxycycline 40 mg) Capsules in Healthy Human Subjects Under FedConditions

The aim of the study was to compare the bioavailability of Doxycycline“drug coated” Capsules 36-mg and Doxycycline “drug coated” Capsules38-mg, relative to the reference product, the FDA approved 40-mg Oracea®capsule under fed conditions.

The bioequivalence study showed that the bioavailability of Doxycycline“drug coated” Capsules 36-mg and Doxycycline “drug coated” Capsules38-mg, are bioequivalent to the reference product, the FDA approved40-mg Oracea® capsule under fed conditions.

Study Objectives

Pharmacokinetics objectives: Comparison of the rate and extent ofabsorption and exposure of two doses of doxycycline monohydrate capsules(Test products) with Oracea® 40-mg capsules (Reference product) inhealthy subjects in order to determine bioequivalence under fedconditions.

Safety objectives: tolerability and safety of single oraladministrations of three dose levels of doxycycline capsules in healthysubjects under fed conditions.

Study Design

The study was conducted according to an open-label, randomized,three-period crossover design with a wash-out phase of 7 days minimum.Subjects received a single dose of the test formulations (Treatment A,36-mg capsule and treatment B, 38-mg capsule) and the reference(Treatment C, 40-mg capsule) products 30 minutes after a 1000 calorie,high-fat and high-protein meal.

Summary of Study Results Study Characteristics

The study was carried out in 44 healthy volunteers; one subject wasexcluded from the bioequivalence assessment due to a major deviation(subject vomited within a six-hour period following drugadministration).

Bioequivalence Assessment:

Blood samples for pharmacokinetic profiling were taken up to 96 hourspost-dose, and doxycycline plasma concentrations were determined with avalidated LC-MS/MS method, LOQ: 15 ng/mL.

The pharmacokinetic (PK) parameters for doxycycline were determinedusing a standard non-compartmental method, and C_(max), AUC_(0-t) (witht being the last quantifiable point), AUC_(0-inf) were analyzedstatistically using log-transformed data. Bioequivalence was assessed byexamining the 90% confidence intervals for the ratio of the testformulation mean relative to the reference formulation mean.

The mean and standard deviations for the PK parameters are provided inTables 5 and 6 below, and the mean plasma profiles in normal scale andlog scale are shown in FIGS. 3 and 4. The statistical analyses of PKresults are summarized in Tables 7 and 8.

36-Mg Doxycycline Capsule Vs. 40 mg Oracea Capsule:

The calculated 90% confidence intervals of the mean test/referenceratios (expressed in percentages) of AUC_(0-t), AUC_(0-inf), C_(max)were [93.57; 100.18], [92.28; 100.34] and [96.51; 104.85], respectively.Of note, the FDA guidance (Bioavailability and Bioequivalence Studiesfor Orally Administered Drug Products—General Considerations—March 2003)stated that to be inside the acceptance interval, values should be atleast 80.00 and no more than 125.00. Therefore, all tested PK parameters(AUC_(0-t), AUC_(0-inf), C_(max)) were strictly within thebioequivalence range of 80.00-125.00%.

38-Mg Doxycycline Capsule Vs. 40 mg Oracea Capsule:

The calculated 90% confidence intervals of the mean test/referenceratios of, AUC_(0-t), AUC_(0-inf) C_(max) were [97.50; 105.57], [100.92;111.09] and [97.55; 108.99], respectively. Therefore, all tested PKparameters (AUC_(0-t), AUC_(0-inf), C_(max)) were strictly within thebioequivalence range of 80.00-125.00%.

In conclusion, Doxycycline 36-mg capsule and 40-mg Oracea® capsule andDoxycycline 38-mg capsule and 40-mg Oracea® under fed conditions areconsidered bioequivalent with respect to C_(max), AUC_(0-t) andAUC_(0-inf) using the 80-125% FDA required criteria.

Food Effect:

Concomitant administration of a 1000 calorie, high-fat, high-proteinmeal with doxycycline 36-mg capsule resulted in a decrease of thesystemic exposure to doxycycline: the rate (C_(max)) and extent ofabsorption (AUC_(0-t)) are 42% and 17% lower than that observed in thestudy under fasting conditions (study # RD.06.SPR.18210). For theformulation containing 38-mg of doxycycline, the food effect was similarand the decrease in the C_(max) and AUC_(0-t) was by 38% and 13%respectively. However, the food effect on doxycycline pharmacokineticsis well known and the observed reductions in C_(max) and AUC_(0-t) inthis study for Oracea (44% and 23%, respectively) were consistent withthe drug approval package (NDA 50-805) for Oracea® (under fed conditionsC_(max) and AUC_(0-t) decreased by 45% and 22%, respectively).

TABLE 5 Overall pharmacokinetics parameters (Arithmetic mean ± SD) 36-mgDoxycycline 40-mg Oracea ® Capsule Capsule Parameters N (TEST) N(REFERENCE) AUC_(0-t) 43 43 (ng · hr/mL) Mean ± SD 5312 ± 1626 5491 ±1336 Min-Max 2264-8922 3673-8753 CV (%) 31% 24% AUC_(0-inf) 28 32 (ng ·hr/mL) Mean ± SD 5769 ± 1542 5793 ± 1146 Min-Max 4022-10089 4391-8794 CV(%) 27% 20% C_(max) (ng/mL) 43 43 Mean ± SD 291 ± 94  288 ± 76  Min-Max155-524 168-477 CV (%) 32% 26% AUC: area under the concentration timecurve; C_(max): observed peak drug concentration; Note: AUC_(0-inf) werecalculated only for profiles meeting acceptance criteria in linearregression.

TABLE 6 Overall pharmacokinetics parameters (Arithmetic mean ± SD) 38-mgDoxycycline 40-mg Oracea ® Capsule Capsule Parameters N (TEST) N(REFERENCE) AUC_(0-t) 43 43 (ng · hr/mL) Mean ± SD 5682 ± 1793 5491 ±1336 Min-Max 3680-10955 3673-8753 CV (%) 32% 24% AUC_(0-inf) 35 32 (ng ·hr/mL) Mean ± SD 6241 ± 1681 5793 ± 1146 Min-Max  4148-10846 4391-8794CV (%) 27% 20% C_(max) (ng/mL) 43 43 Mean ± SD 304 ± 117 288 ± 76 Min-Max 143-750 168-477 CV (%) 39% 26% AUC: area under the concentrationtime curve; C_(max): observed peak drug concentration; Note: AUC_(0-inf)were calculated only for profiles meeting acceptance criteria in linearregression.

TABLE 7 Summary of statistical analysis of pharmacokinetic data (36-mgvs 40-mg) Least Square Mean Geometric Mean^(a) 90% Confidence Interval36 mg 40 mg 36 mg 40 mg Lower Upper Doxycycline Oracea ® DoxycyclineOracea ® Ratio Limit Limit Capsule Capsule Capsule Capsule (%) (%) (%)AUC_(0-t) 8.545 8.577 5140.31 5309.42 96.81 93.57 100.18 (ng · hr/mL)AUC_(0-inf) 8.638 8.676 5640.28 5861.40 96.23 92.28 100.34 (ng · hr/mL)C_(max) 5.635 5.629 280.06 278.40 100.59 96.51 104.85 (ng/mL)^(a)Geometric means are based on least square means or ln-transformedvalues

TABLE 8 Summary of statistical analysis of pharmacokinetic data (38-mgvs 40-mg) Least Square Mean Geometric Mean^(a) 90% Confidence Interval38 mg 40 mg 38 mg 40 mg Lower Upper Doxycycline Oracea ® DoxycyclineOracea ® Ratio Limit Limit Capsule Capsule Capsule Capsule (%) (%) (%)AUC_(0-t) 8.591 8.577 5385.53 5308.36 101.45 97.50 105.57 (ng · hr/mL)AUC_(0-inf) 8.704 8.646 6024.67 5689.95 105.88 100.92 111.09 (ng ·hr/mL) C_(max) 5.658 5.627 286.45 277.81 103.11 97.55 108.99 (ng/mL)^(a)Geometric means are based on least square means of ln-transformedvalues

Safety

No Serious Adverse Events (SAEs) were reported.

There were no discontinuations due to AEs.

32 Adverse Events (AEs) were reported for this study, of which 13 wererelated to study drug. These related AEs occurred in 10 subjects, andwere mostly related to nervous system complaints (headache) andgastrointestinal system disturbances (nausea and vomiting). All relatedAEs were transient, and mild to moderate in severity; they were alsoconsistent with the known safety/tolerability profile of doxycycline.

A summary of Treatment Emergent Adverse Events (TEAEs) is provided inTable 9.

TABLE 9 Overall Summary of Treatment Emergent Adverse Events [SafetyPopulation] 36 mg 38 mg 40 mg Doxycycline Doxycycline Oracea ® CapsuleCapsule Capsule (N = 44) (N = 44) (N = 44) n (%) E n (%) E n (%) E AnyTEAEs 8 (18.2%) 12 10 (22.7%) 15 3 (6.8%) 5 Any Drug- 4 (9.1%) 0 5(11.4%) 7 1 (2.3%) 1 Related TEAEs Any SAEs 0 (0.0%) 0 0 (0.0%) 0 0(0.0%) 0 Any Drug- 0 (0.0%) 0 0 (0.0%) 0 0 (0.0%) 0 Related SAEs AnyDeaths 0 (0.0%) 0 0 (0.0%) 0 0 (0.0%) 0 Any Drug- 0 (0.0%) 0 0 (0.0%) 00 (0.0%) 0 Related Deaths Any TEAEs 0 (0.0%) 0 0 (0.0%) 0 0 (0.0%) 0Leading to Withdrawal Any SAEs 0 (0.0%) 0 0 (0.0%) 0 0 (0.0%) 0Resulting in Early Termination Note: N: Number of Subjects in theSpecified Group n: Number of Subjects reporting TEAEs %: Percentagebased on N E: Number of TEAEs

No pregnancies were reported.

Given the study design (single-dose, three-way crossover) and smallsample size, it is self-evident that any safety findings are not fullyrepresentative of use in the recommended clinical setting in a largerpopulation. Overall however, no new safety or tolerability issues wereraised that are not addressed in the approved labelling for Oracea®capsules.

CONCLUSION

The present bioequivalence study showed that the bioavailability ofDoxycycline “drug coated” Capsules 36-mg and Doxycycline “drug coated”Capsules 38-mg, are bioequivalent to the reference product, the FDAapproved 40-mg Oracea® capsule under fed conditions.

Example 3 Batch Formula

The batch formula presented in the Table 10 shows an industrial batchsize of 125 000 capsules of the present invention.

TABLE 10 Doxycycline coated capsules, 36 mg and 38 mg - Batch formulaDoxycycline coated Doxycycline coated capsules, 36 mg capsules, 38 mgUnit Unit formula formula (350.8 Quantity (353.5 Quantity Ingredient mg)per batch mg) per batch Inactive capsules 263.5 mg 32 938 g 263.5 mg 32938 g Seal coating (13.3 mg) Methacrylic acid 11.1 mg 4625 g 11.1 mg4625 g copolymer dispersion (Eudragit ® L3OD 55) Triethyl citrate 2.2 mg275 g 2.2 mg 275 g Drug layer delayed release (5.39 mg for 36 mgcapsules and 8.09 for 38 mg capsules) Hydroxypropyl 1.33 mg 166 g 2.00mg 249 g methylcellulose (Methocel E6LVCI) Simethicone 0.06 mg 25 g 0.09mg 38 g emulsion (Simethicone 30% emulsion) Doxycycline 4 mg 500 g* 6 mg750 g* (as monohydrate, micronized) Enteric coating (18.5 mg)Methacrylic acid 15.4 mg 6417 g 15.4 mg 6417 g copolymer dispersion(Eudragit ® L3OD 55) Triethyl citrate 3.1 mg 391 g 3.1 mg 391 g Druglayer immediate release (43.1 mg) Hydroxypropyl 10.6 mg 1325 g 10.6 mg1325 g methylcellulose (Methocel E6LVCI) Simethicone 0.49 mg 204 g 0.49mg 204 g emulsion (Simethicone 30% emulsion) Doxycycline (asmonohydrate, 32 mg 4000 g* 32 mg 4000 g* micronized) Color coating (7mg) Opadry ® blue, 7 mg 875 g 7 mg 875 g Y-S-4258 Printing Opacode ® QSQS QS QS Monogramming Ink S-1-18089 White (″) The required quantity ofdoxycycline monohydrate to be used (Q) is calculated taking into accountthe water content (M %) and the purity of the anhydrous drug substance(P %).Example: for 500 g of doxycycline monohydrate required, the quantity tobe used (Q) is as follows:

Q(g)=[500 g*100*100]/[P*(100−M)]

For feasibility and demonstration batches, a 2% excess drug coatingsuspension (for delayed and immediate release layers) is prepared tocompensate the processing loss and therefore, not added in the batchformula. From registration batches, the excess drug coating suspensionwill be of 3% based on feasibility and demonstration batches results.

Example 4

TABLE 11 Doxycycline coated capsules, 36 mg and 38 mg- Qualitative andquantitative compositions Doxycycline Doxycycline Capsules, Capsules, 36mg 38 mg Unit formula Unit formula Ingredient (350.8 mg) (353.5 mg)Inactive capsules 1 1 (Approximately (Approximately 263.5 mg) 263.5 mg)Seal coating (13.3 mg) Methacrylic acid copolymer dispersion 11.1 mg11.1 mg (Eudragit ® L300 55) Triethyl citrate 2.2 mg 2.2 mg Drug layerdelayed release (5.39 mg for 36 mg capsules and 8.09 mg for 38 mgcapsules) Hydroxypropyl methylcellulose 1.33 mg 2.00 mg (MethocelE6LVC)s) Simethicone emulsion 0.06 mg 0.09 mg (Simethicone 30% emulsion)Doxycycline 4 mg 6 mg Enteric coating (18.5 mg) Methacrylic acidcopolymer dispersion 15.4 mg 15.4 mg (Eudragit ® L3OD 55) Triethylcitrate 3.1 mg 3.1 mg Drug layer immediate release (43.1 mg)Hydroxypropyl methylcellulose 10.6 mg 10.6 mg (Methocel E6LVC)s)Simethicone emulsion 0.49 mg 0.49 mg (Simethicone 30% emulsion)Doxycycline 32 mg 32 mg Color coating (7 mg) Opadry ® blue, Y-S-4258 7mg 7 mg Printing Opacode ® Monogramming Ink QS QS S-1-18089 White

Physicochemical and Biological Properties

The unit form doses of the present invention exhibit the samedissolution characteristics as Oracea® Capsules, 40 mg (30 mg immediaterelease and 10 mg delayed release) marketed by Galderma, and have abioequivalent profile.

Dissolution Profiles

A pilot-scale batch (Batch PB714) of Doxycycline coated capsules, 40 mg(30 mg IR/10 mg DR) was manufactured, i.e., an outside coated capsulecontaining 40 mg doxycycline distributed in two layers: a pH dependentlayer containing 10 mg doxycycline for delayed release and a layercontaining 30 mg doxycycline for immediate release. This batch was usedin PK study BE09/040-BE091041, to assess bioequivalence comparatively toOracea capsules (Batch 0805129A).

The dissolution test used for the development is the one that isspecified in the FDA/OGD website for Doxycycline capsules (delayedrelease) 40 mg, as described in Table 12.

TABLE 12 Dissolution conditions Dissolution media Buffer A, 40 mLconcentrated HCl in 6000 mL pH 1.1 of water Buffer B 54.4 g potassiumphosphate dibasic + 8.1 g NaOH pellets in 2000 mL of water Buffer C 40 gNaOH pellets in 500 mL of water Medium volume 750 mL Buffer A (pH 1.1)950 mL Buffer B + Buffer C (pH 6.0) Medium 37° C. + 0.5° C. temperatureApparatus USP Apparatus ll (paddles) Paddle rotation 75 RPM Dissolutiontime 2 h at pH 1.1 (Buffer A) 2 h at pH 6.0 (after addition of BufferB + Buffer C) Sampling times 30, 60, 120, 150, 180 and 240 minutes Assaymethod UV 268 nm Cell path length 0.5 cm Standard solution 40 mgdoxycycline HCl in 50 mL Buffer A, diluted to 1/25 in Buffer A Finalconcentration: 0.032 mg/mL doxycycline HCl

The dissolution profiles of this formulation and Oracea capsulespresented in FIG. 6 indicate that both formulations present a similarprofile.

The dissolution profiles of two demonstration batches (Batch CD000911,capsules coated with 36 mg doxycycline drug substance: 4 mg in DRlayer/32 mg in IR layer and Batch CD001111, capsules coated with 38 mgdoxycycline drug substance: 6 mg in DR layer/32 mg in IR layer), testedaccording to the conditions described in Table 12 above are presented inFIG. 7 and FIG. 8.

Dissolution profiles performed as in-process controls on the capsulescoated with only the delayed release layer (i.e. with 4 mg and 6 mgdoxycycline for the 36 mg and 38 mg strengths, respectively) at the endof step 4 of the manufacturing process, before application of theimmediate release drug coating layer are presented in FIG. 7.

Dissolution profiles performed on the finished drug product, i.e. thecapsules coated with 36 mg and 38 mg doxycycline (4 mg DR/32 mg IR. and6 mg DR/32 mg IR, respectively) are presented in FIG. 8.

The dissolution profiles indicate that approximately 80% to 90% of thetotal amount of doxycycline is released in 30 minutes at pH 1.1, i.e.the totality of the drug substance present in the immediate releaselayer.

After 240 minutes (120 minutes at pH 1.1 then 120 minutes at pH 6.0),approximately 90% of the total amount of doxycycline are dissolved,corresponding to the subsequent dissolution of the delayed releaselayers.

The dissolution profiles, the formulation and the manufacturing processof the capsules, confirm that this dosage form presents both immediaterelease and delayed release properties.

These formulations (capsules coated with 36 mg and 38 mg doxycycline (4mg DR/32 mg IR, and 6 mg DR/32 mg IR, respectively)) are those intendedto be tested in pivotal PK studies. They correspond to the industrialmanufacturing process and batch size (125 000 capsules described above).

Pharmacokinetic Data

Doxycycline coated capsules, 40 mg (30 mg IR/10 mg/DR), were tested in apharmacokinetic study under fasting and fed conditions, aimed to assessbioequivalence comparatively to Oracea® capsules.

The formulation was found bioequivalent to Oracea capsules, 40 mg (seeFIG. 9); moreover, the results obtained suggested that bioequivalencecould be fine-tuned with slightly different proportions of doxycycline.

A second study under fasting and fed conditions was then undertaken withtwo formulations respectively coated with 36 mg doxycycline (4 mgdelayed release and 32 mg immediate release and) and 37.5 mg doxycycline(7.5 mg delayed release and 30 mg immediate release) comparatively toOracea® Capsules, 40 mg. The formulation at 36 mg was foundbioequivalent both under fasting and fed conditions while formulation at37.5 mg was bioequivalent under fed conditions only (see FIG. 10 andFIG. 11).

Dissolution and PK Data Relationship

The dissolution profiles of the different formulations clearly displaythe presence of an immediate release layer and a delayed release layerin the doxycycline coated capsules. The pharmacokinetic data display thepart played by the ratio IR/DR in the formulation: the IR layer enhancesC._(a)x and the DR layer enhances AUC (terminal slope).

Thus, while there have been described what are presently believed to bethe preferred embodiments of the present invention, those skilled in theart will realize that other and further embodiments can be made withoutdeparting from the spirit of the invention, and it is intended toinclude all such further modifications and changes as come within thetrue scope of the claims set forth herein.

We claim:
 1. A pharmaceutical composition in unit dose form for orallydelivering doxycycline to a human, the pharmaceutical compositioncomprising: a capsule, wherein the capsule is coated with a delayedrelease layer; wherein the delayed release layer comprises about 4 to 6mg of doxycycline, or a pharmaceutically acceptable salt thereof, and abinding agent, and wherein the delayed release layer is coated with anenteric coating; wherein the enteric coating dissolves at pH of about 5to 6, and wherein the enteric coating is coated with an immediaterelease layer; wherein the immediate release layer comprises about 32 mgof doxycycline, or a pharmaceutically acceptable salt thereof, and abinding agent, wherein the relative mean C_(max) of the pharmaceuticalcomposition is within 80.00% to 125.00% of a C_(max) value of 510±220.7ng/mL, after administration of a single dose of the pharmaceuticalcomposition to humans in a fasting state; and wherein the relative meanAUC_((0-∞)) of the pharmaceutical composition is within 80.00% to125.00% of a AUC_((0-∞)) value of 9227±3212.8 ng·hr/mL, afteradministration of a single dose of the pharmaceutical composition tohumans in a fasting state.
 2. The pharmaceutical composition of claim 1,wherein the relative mean C_(max) of the pharmaceutical composition atsteady state is within 80.00% to 125.00% of a C_(max) value of 600±194.2ng/mL after administration to humans in a fasting state, and wherein therelative mean AUC_((0-t)) of the pharmaceutical composition at steadystate is within 80.00% to 125.00% of a AUC_((0-t)) value of 7543±2443.9ng·hr/mL, after administration to humans in a fasting state.
 3. Thepharmaceutical composition of claim 1 wherein the delayed release layercomprises about 4 mg of doxycycline monohydrate.
 4. The pharmaceuticalcomposition of claim 1 wherein the delayed release layer comprises about6 mg of doxycycline monohydrate.
 5. The pharmaceutical composition ofclaim 1 wherein the doxycycline salt is micronized.
 6. Thepharmaceutical composition of claim 5 wherein the micronized doxycyclinesalt has less than about 10 μm distribution for at least about 90% ofparticles.
 7. The pharmaceutical composition of claim 1 furthercomprising a seal coating between the capsule and the delayed releasecoating.
 8. The pharmaceutical composition of claim 7 wherein the sealcoating comprises a pH dependent ingredient, wherein the pH dependentingredient dissolves at pH of about 5 to
 6. 9. The pharmaceuticalcomposition of claim 1 wherein the capsule contains about 200 mg toabout 260 mg of at least one inert ingredient.
 10. The pharmaceuticalcomposition of claim 9 wherein the capsule does not contain an activeingredient.
 11. The pharmaceutical composition of claim 1 wherein theratio of the binding agent to the doxycycline monohydrate in the delayedrelease layer is about 1:3.
 12. The pharmaceutical composition of claim1 wherein the ratio of the binding agent to the doxycycline monohydratein the immediate release layer is about 1:3.
 13. The pharmaceuticalcomposition of claim 1 wherein the binding agent is hydroxypropylmethylcellulose (HPMC), hydroxyethyl cellulose (HEC), hydroxypropylcellulose (HPC), carboxy methyl cellulose (CMC), or mixtures thereof.14. The pharmaceutical composition of claim 1, having a dissolutionprofile such that about 80% to about 90% of the doxycycline monohydrateis dissolved after about 30 minutes at pH of about 1.1.
 15. Thepharmaceutical composition of claim 1, having a dissolution profile suchthat about 90% of the doxycycline monohydrate is dissolved after about120 minutes at pH of about 1.1 and 120 minutes at pH of about 6.0. 16.The pharmaceutical composition of claim 1, having a dissolution profileas shown in FIG.
 8. 17. The pharmaceutical composition of claim 1,wherein the capsule is hydroxypropyl methylcellulose.
 18. Thepharmaceutical composition of claim 1, wherein the 90% ConfidenceInterval of the relative mean C_(max), AUC_((0-t)) and AUC_((0-∞)) ofthe pharmaceutical composition to a reference formulation is within80.00% to 125.00% upon administration to humans in a fasting state,wherein the reference formulation is a gelatin capsule filled with beadsof doxycycline monohydrate, wherein the beads consist of 30 mg ofimmediate release beads and 10 mg of delayed release beads.
 19. A methodfor treating papules and pustules of rosacea in a human in need thereof,comprising orally administering a once a day dose of a pharmaceuticalcomposition to the human, the pharmaceutical composition comprising: acapsule, wherein the capsule is coated with a delayed release layer;wherein the delayed release layer comprises about 4 to 6 mg ofdoxycycline, or a pharmaceutically acceptable salt thereof, and abinding agent, and wherein the delayed release layer is coated with anenteric coating; wherein the enteric coating dissolves at pH of about 5to 6, and wherein the enteric coating is coated with an immediaterelease layer; wherein the immediate release layer comprises about 32 mgof doxycycline, or a pharmaceutically acceptable salt thereof, and abinding agent, wherein the relative mean C_(max) of the pharmaceuticalcomposition is within 80.00% to 125.00% of a C_(max) value of 510±220.7ng/mL, after administration of a single dose of the pharmaceuticalcomposition to humans in a fasting state; and wherein the relative meanAUC_((0-∞)) of the pharmaceutical composition is within 80.00% to125.00% of a AUC_((0-∞)) value of 9227±3212.8 ng·hr/mL, afteradministration of a single dose of the pharmaceutical composition tohumans in a fasting state.
 20. The method of claim 19, wherein therelative mean C_(max) of the pharmaceutical composition at steady stateis within 80.00% to 125.00% of a C_(max) value of 600±194.2 ng/mL afteradministration to humans in a fasting state, and wherein the relativemean AUC_((0-t)) of the pharmaceutical composition at steady state iswithin 80.00% to 125.00% of a AUC_((0-t)) value of 7543±2443.9 ng·hr/mL,after administration to humans in a fasting state.
 21. The method ofclaim 19, wherein the pharmaceutical composition further comprises aseal coating between the capsule and the delayed release coating,wherein the seal coating comprises a pH dependent ingredient, whereinthe pH dependent ingredient dissolves at pH of about 5 to
 6. 22. Themethod of claim 19 wherein the delayed release layer comprises about 4mg of doxycycline monohydrate.
 23. The method of claim 19 wherein thedelayed release layer comprises about 6 mg of doxycycline monohydrate.24. The method of claim 19, wherein the pharmaceutical composition has adissolution profile such that about 80% to about 90% of the doxycyclinesalt is dissolved after about 30 minutes at pH of about 1.1.
 25. Themethod of claim 19, wherein the pharmaceutical composition has adissolution profile such that about 90% of the doxycycline salt isdissolved after about 120 minutes at pH of about 1.1 and 120 minutes atpH of about 6.0.
 26. The method of claim 19, wherein pharmaceuticalcomposition has a dissolution profile as shown in FIG.
 8. 27. Apharmaceutical composition in unit dose form for orally deliveringdoxycycline to a human, the pharmaceutical composition comprising: acapsule, wherein the capsule is coated with a delayed release layer;wherein the delayed release layer comprises about 4 to 6 mg ofdoxycycline, or a pharmaceutically acceptable salt thereof, and abinding agent, and wherein the delayed release layer is coated with anenteric coating; wherein the enteric coating dissolves at pH of about 5to 6, and wherein the enteric coating is coated with an immediaterelease layer; wherein the immediate release layer comprises about 32 mgof doxycycline, or a pharmaceutically acceptable salt thereof; andwherein after administration of a single dose to humans in a fastingstate, the C_(max) has a range of about 230 ng/mL to about 1120 ng/mL,and the AUC_((0-∞)) has a range of about 4800 ng·hr/mL to about 15,600ng·hr/mL.
 28. The pharmaceutical composition of claim 27 wherein atsteady state the C_(max), has a range of about 325 ng/mL to about 1000ng/mL, and the AUC_((0-∞)) has a range of about 3500 ng·hr/mL to about12,500 ng·hr/mL.